KR20220020951A - CDK kinase inhibitors - Google Patents

Abstract

Disclosed herein are compounds of general formula (I) and salts thereof which can be used as CDK kinase (in particular CDK4/6 kinase) inhibitors, all modifications being defined in the present text. The compounds can be used for the treatment or prevention of diseases such as cancer. The application further relates to a pharmaceutical composition comprising a compound of formula (I).

Description

CDK kinase inhibitors

The present invention relates to compounds which inhibit the activity of CDK kinases and to the use of said compounds in the manufacture of a medicament for the treatment and/or prophylaxis of cancer-related diseases mediated by CDK kinases.

Cyclin-dependent kinases (CDKs) are a group of serine/threonine protein kinases that are closely related to important cellular processes such as cell cycle or transcriptional regulation. Research has shown that instead of exerting kinase activity by itself, CDKs exert protein kinase activity by binding to cyclins and forming specific CDK-cyclin complexes, thereby promoting cell cycle phase transitions, initiating DNA synthesis, It has been suggested that it has a function such as regulating cellular transcription.

The current CDK family consists of 13 members (CDK1 to CDK13). They are divided into two main categories according to their intracellular functions: CDKs that regulate the cell cycle (CDK1, CDK2, CDK4, CDK6, etc.) and CDKs that regulate transcription (CDK7, CDK9, etc.). There are 11 subtypes of cyclins named after A-I, k, and T. Its expression is transcriptionally regulated and fluctuates regularly during the cell cycle. CDK4/6 plays an irreplaceable role in the CDK subtypes involved in cell cycle regulation. Cancer-associated cell cycle mutations are primarily present during the G1 phase and G1/S transition. CDK4/6 binds to cyclin D to form a complex with kinase activity, which releases the bound transcription factor E2F through phosphorylation of oncostatin Rb, initiates transcription of genes associated with S phase, and allows cells to pass and cause a shift from the G1 phase to the S phase. Specific activation of CDK4/6 is closely related to the proliferation of specific tumors. There is an aberrant cyclin D-CDK4/6-INK4-Rb pathway in about 80% of human tumors. Alterations in this pathway may accelerate the progression of the G1 phase, thereby accelerating the proliferation and survival benefits of tumor cells. Therefore, it is a therapeutic strategy that interferes with the pathway and CDK4/6 becomes one of the antitumor targets.

To date, more than 50 CDK inhibitors have been reported, some of which have potential antitumor activity. Some broad-spectrum CDK inhibitors have been developed as antitumor drugs, and some are in preclinical or clinical trials. New CDK inhibitors are constantly being developed. Flavopyridol, now also known as the CDK inhibitor L86-8275 or HMR1275, represents the first generation of CDK inhibitors. It did not enter phase 3 clinical trials due to unclear efficacy and high toxicity.

Recently, some pharmaceutical companies, such as Pfizer, Eli Lilly, and Novartis, have reported successive series of CDK inhibitors with good selectivity and are conducting clinical trials. Of particular importance are palbociclib (PD-0332991) developed by Pfizer, avemaciclib (LY2835219) developed by Eli Lilly, and ribociclib (LEE011) developed by Novartis.

Palbociclib is a highly specific CDK4- (IC ₅₀ =0.011 μmol/L) and CDK6- (IC ₅₀ =0.016 μmol/L) selective inhibitor developed by Pfizer (see Documents 1 and 2). Palbociclib is inactive against 36 protein kinases, including the other CDK tyrosine/serine and threonine kinases. In February 2015, palbociclib was approved by the US FDA for the treatment of patients with estrogen receptor-positive breast cancer. The successful listing of these compounds once again caused a wave of CDK inhibitor development.

Abemaciclib is an effective oral cyclin dependent kinase (CDK) inhibitor targeting the CDK4 (cyclin D1) and CDK6 (cyclin D3) cell cycle pathways and has potential antitumor activity. Abemaciclib specifically inhibits CDK4/6 to inhibit phosphorylation of retinoblastoma (Rb) protein in the early G1 phase. Inhibition of Rb phosphorylation prevents CDK-mediated G1-S transition, resulting in cell cycle arrest at G1 phase, inhibiting DNA synthesis and inhibiting cancer cell growth. In March 2017, Eli Eley published a successful phase 3 clinical study of abemaciclib in the treatment of breast cancer.

Ribociclib is another orally effective CDK4 and CDK6 selective inhibitor (IC ₅₀ is 10 and 39 nmol/L, respectively) (see Documents 3 and 4). As expected, ribociclib inhibits Rb phosphorylation, induces G0/G1 phase arrest, and induces senescence of tumor cells (including melanoma, breast cancer, liposarcoma, and gliocytoma with B-Raf or N-Ras mutations). On March 14, 2017, Ribociclib was approved by the US FDA for combination therapy with an aromatase inhibitor for the treatment of hormone receptor positive and human epidermal growth factor receptor 2 negative postmenopausal women.

Currently, there is an increasing number of studies around the world focusing on this goal. Small molecule inhibitors (especially CDK4/6 kinase inhibitors) that target CDKs have high development value. There is a lot of room for development, and it has great significance in exploring new antitumor drugs in this field.

prior art literature

Patent Document 1: WO2003062236A1;

Patent Document 2: WO2005005426A1;

Patent Document 3: WO2011101409A1;

Patent Document 4: CN103788100A.

It is an object of the present invention to provide kinase inhibitor compounds for CDKs.

The present invention provides a compound of formula (I): or a pharmaceutically acceptable salt thereof:

during the meal,

Q is an optionally substituted 6- to 18-membered arylene group or an optionally substituted 5- to 18-membered heteroarylene group, wherein when substituted, the substituents are halo, _hydroxy , C _1-6 alkyl, selected from the group consisting of C _1-6 alkoxy, and halo _- C _{1-6 alkyl} ;

R ₁ is an optionally substituted 3- to 8-membered heterocyclyl group, an optionally substituted 6- to 14-membered fused heterocyclyl group, or an optionally substituted 6- to 12-membered spiro heterocyclyl group wherein, when substituted, the substituent is selected from the group consisting of halo, hydroxy, C _1-6 alkyl, C _1-6 alkoxy, and halo-C _1-6 alkyl;

R ₂ is H, halo, optionally substituted 3- to 10-membered cycloalkenyl group, optionally substituted 3- to 10-membered heterocycloalkenyl group, optionally substituted 3- to 10-membered cycloalkyl group, an optionally substituted 3- to 10-membered heterocycloalkyl group, an optionally substituted 6- to 18-membered aryl group, or an optionally substituted 5- to 18-membered heteroaryl group, wherein substituted , the substituents are selected from the group consisting of halo, _hydroxy , C _1-6 alkyl, C _1-6 alkoxy, halo _- C _{1-6 alkyl} , C _1-6 alkoxy _- C _1-6 alkoxy, and oxo;

R ₃ is H, CN, —C(=O)—NR ₄ R ₅ , an optionally substituted 6- to 18-membered aryl group, an optionally substituted 5- to 18-membered heteroaryl group, or optionally substituted a 5- to 8-membered lactam group wherein, when substituted, the substituent is selected from the group consisting of halo, _hydroxy , C _{1-6 alkyl} , C _1-6 alkoxy, and halo-C _1-6 alkyl;

R ₄ and R ₅ are each independently methyl or ethyl; And

when R ₃ is —C(=O)—NR ₄ R ₅ , then R ₂ is an optionally substituted 3- to 10-membered cycloalkenyl group, an optionally substituted 3- to 10-membered heterocycloalkenyl group , an optionally substituted 3- to 10-membered heterocycloalkyl group linked at the N atom to a non-R ₂ structure of Formula (I), or an optionally substituted 6- to 18-membered aryl group, wherein substituted , the substituent is selected from the group consisting of halo, _hydroxy , C _1-6 alkyl, C _1-6 alkoxy, halo _- C _{1-6 alkyl} , C _1-6 alkoxy _- C _1-6 alkoxy, and oxo.

In a preferred embodiment,

Q is optionally substituted phenylene or optionally substituted pyridinylene, wherein when substituted, the substituents are halo, _hydroxy , C _{1-6 alkyl} , C _1-6 alkoxy, and halo _- C _1-6 alkyl. is selected from the group consisting of;

R ₁ is an optionally substituted 3- to 8-membered heterocyclyl group, an optionally substituted 6- to 14-membered fused heterocyclyl group, or an optionally substituted 6- to 12-membered spiro heterocyclyl group wherein when substituted, the substituent is selected from C _{1-6 alkyl} ;

R ₂ is a halogen atom, optionally substituted cyclopentenyl, optionally substituted cyclohexenyl, optionally substituted cyclopentyl, optionally substituted cyclohexyl, optionally substituted oxacyclohexenyl, optionally substituted aza cyclohexenyl, optionally substituted oxolanyl, optionally substituted azacyclopentyl, optionally substituted oxacyclohexyl, optionally substituted azacyclohexyl, optionally substituted phenyl, optionally substituted naphthyl, optionally is selected from the group consisting of pyridinyl substituted with , optionally substituted thienyl, optionally substituted pyrazolyl, optionally substituted oxazolyl, optionally substituted isoxazolyl, and optionally substituted quinolyl, wherein When substituted, the substituent is selected from the group consisting of halo, _hydroxy , C _1-6 alkyl, C _1-6 alkoxy, halo _- C _1-6 alkyl, and oxo;

또는

이고, 여기서, 치환된 경우, 치환기는 할로, 하이드록시, C_{1- 6}알킬, C_{1- 6}알콕시 및 할로-C_{1- 6}알킬로 이루어진 군으로부터 선택되고; 그리고R ₃ is H, -CN, -C(=O)-NR ₄ R ₅ , optionally substituted phenyl, naphthyl, pyrazolyl, pyridinyl, thienyl, oxazolyl, isoxazolyl, pyrimidinyl, imine Dazolyl, pyrrolyl,

or

wherein, when substituted, the substituent is selected from the group consisting of halo, _hydroxy , C _{1-6 alkyl} , C _1-6 alkoxy and halo _- C _1-6 alkyl; And

R ₄ and R ₅ are both methyl.

이다.In another preferred embodiment, Q is phenylene or

am.

In another preferred embodiment, R ₁ is a group optionally substituted as follows, wherein when substituted, the substituent is selected from C _{1-6 alkyl} :

In another preferred embodiment, C _1-6 alkyl is methyl or ethyl.

In another preferred embodiment, R ₂ is halo or a group optionally substituted as follows, wherein when substituted, the substituent is selected from C _{1-6 alkyl} :

In another aspect of the present invention, each exemplary compound of Table 1 or a pharmaceutically acceptable salt thereof is provided.

In another aspect of the invention, the compounds of the invention are used to inhibit the activity of a CDK kinase; In particular, the compounds of the present invention are used to inhibit the activity of CDK4/6 kinases.

In another aspect of the present invention, there is provided a pharmaceutical composition comprising an effective amount of the compound of the present invention, and a pharmaceutically acceptable carrier or excipient.

In another embodiment of the present invention, in the preparation of a medicament for the treatment and/or prophylaxis of cancer-related diseases mediated by CDK kinase (especially CDK4/6 kinase), the compound of the present invention or a pharmaceutically acceptable salt thereof Use is provided, wherein the cancer-related disease is brain tumor, lung cancer, squamous cell carcinoma, bladder cancer, stomach cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, rectal cancer, liver cancer, kidney cancer, Esophageal adenocarcinoma, esophageal squamous cell carcinoma, prostate cancer, female reproductive tract cancer, carcinoma in situ, lymphoma, neurofibroma, thyroid cancer, bone cancer, skin cancer, brain cancer, colon cancer, testicular cancer, gastrointestinal stromal tumor, prostate tumor, mast cell tumor, multiple myeloma , melanoma, glioma, or sarcoma.

The compounds of the present invention can inhibit the activity of CDK kinases (especially CDK4/6 kinases) and can be used in the treatment of cancer.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the claimed subject matter belongs.

"C _{1-6 alkyl} " refers to all possible isomeric forms such as methyl, ethyl, propyl, butyl, pentyl and hexyl, including n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, and the like. refers to an alkyl group having 1 to 6 carbon atoms including “C _{1-6 alkyl} ” includes all sub _- ranges subsumed therein, such as C _1-2 alkyl, C _1-3 alkyl, C _{1-4 alkyl} , C _{1-5 alkyl} , C _{2-5 alkyl} , _{C 3 - 5} alkyl, C _4-5 alkyl, C _3-4 alkyl, _{C 3-5} alkyl, _{and C 4-5 alkyl} .

“Aryl” and “heteroaryl” include monocyclic or fused polycyclic (ie, rings that share adjacent pairs of ring atoms) groups. Examples of “aryl” include, but are not limited to, phenyl, naphthalenyl, phenanthrenyl, anthracenyl, fluorenyl, and indenyl. Examples of "heteroaryl" include:

등.

etc.

The term “6- to 18-membered arylene” as used herein refers to two hydrogen atoms including, for example, “6- to 14-membered arylene,” “6- to 10-membered arylene,” and the like. refers to a divalent group derived from an aromatic hydrocarbon having 6 to 18 carbon atoms by extraction of Examples include, but are not limited to, phenylene, naphthylene, anthrylene, and the like.

The term "5- to 18-membered heteroarylene" as used herein includes, for example, "5- to 14-membered heteroarylene", "5- to 10-membered heteroarylene", and the like. refers to a divalent group derived from a 5- to 18-membered heteroaromatic hydrocarbon by extraction of hydrogen atoms. Examples are pufurylene, thienylene, pyrrolylene, imidazolylene, thiazolylene, pyrazolylene, oxazolylene, isoxazolylene, isothiazolylene, pyridinylene, pyrazinylene, pyridazinylene , pyrimidinylene, etc., preferably pyridinylene, pyrazolylene or thienylene, particularly preferably pyridinylene, and most preferably pyridinylene in which the nitrogen atom is at position 3 of the bonding position of R1; including but not limited to.

The term "3- to 8-membered heterocyclyl" as used herein is, for example, "3- to 7-membered heterocyclyl", "3- to 6-membered heterocyclyl", "4 - to 7-membered heterocyclyl", "4- to 6-membered heterocyclyl", "5- to 7-membered heterocyclyl", "5- to 6-membered heterocyclyl", "6-membered heterocyclyl" and the like. Specific examples include aziridinyl, 2H-aziridinyl, diaziridinyl, 3H-diazirenyl, azetidinyl, 1,4-dioxanyl, 1,3-dioxanyl, 1,3-dioxolanyl , 1,4-dioxacyclohexadienyl, tetrahydrofuryl, dihydropyrrolyl, pyrrolidinyl, imidazolidinyl, 4,5-dihydroimidazolyl, pyrazolidinyl, 4,5-dihydro Pyrazolyl, 2,5-dihydrothienyl, tetrahydrothienyl, 4,5-dihydrothiazolyl, piperidinyl, piperazinyl, morpholinyl, 4,5-dihydrooxazolyl, 4,5 -dihydroisoxazolyl, 2,3-dihydroisoxazolyl, 2H-1,2-oxazinyl, 6H-1,3-oxazinyl, 4H-1,3-thiazinyl, 6H-1,3-thiazinyl , 2H-pyranyl, 2H-pyran-2-onyl, 3,4-dihydro-2H-pyranyl, and the like, preferably “5- to 6-membered heterocyclyl”.

The term “6- to 14-membered fused heterocyclyl” as described herein includes, for example, “6- to 11-membered fused heterocyclyl”, “6- to 10-membered fused heterocyclyl” reyl", "7- to 10-membered fused heterocyclyl", "9- to 10-membered fused heterocyclyl" and the like. Specific examples include, but are not limited to:

The term "6- to 12-membered spiro heterocyclyl" as used herein refers to a ring structure having 6-12 ring atoms containing one or more heteroatoms formed from two or more rings sharing one atom. refers to The heteroatom is selected from N, S, O, CO, SO and/or SO ₂ and the like. This term includes, for example, "6- to 11-membered spiro heterocyclyl", "7- to 11-membered spiro heterocyclyl", "7- to 10-membered spiro heterocyclyl", "7- to 9-membered spiro heterocyclyl", "7- to 8-membered spiro heterocyclyl" and the like. Examples include, but are not limited to:

The term “3- to 10-membered cycloalkenyl” as used herein includes, for example, “3- to 8-membered cycloalkenyl,” “4- to 6-membered cycloalkenyl,” and the like. refers to a cyclic alkenyl group derived from a cyclic monoolefin moiety having 3 to 10 carbon atoms by extraction of one hydrogen atom. Examples include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, cyclononenyl, cyclodecenyl, and the like.

The term "3- to 10-membered heterocycloalkenyl" as used herein, for example, "3- to 8-membered heterocycloalkenyl", "4- to 6-membered heterocycloalkenyl" refers to a heterocycloalkenyl group derived from a 3- to 10-membered cyclic monoolefin moiety containing a heteroatom by extraction of one hydrogen atom including the Examples include, but are not limited to:

The term “3- to 10-membered cycloalkyl” as used herein refers to, for example, one hydrogen atom, including “3- to 8-membered cycloalkyl”, “4- to 6-membered cycloalkyl”, and the like. refers to a cyclic alkyl group derived from a cycloalkane moiety having 3 to 10 carbon atoms by extraction of Examples include, but are not limited to, cyclopropanyl, cyclobutanyl, cyclofentanyl, cyclohexanyl, cycloheptanyl, cyclooctanyl, cyclononanyl, cyclodecanyl, and the like.

The term “3- to 10-membered heterocycloalkyl” as used herein includes, for example, “3- to 8-membered heterocycloalkyl”, “4- to 6-membered heterocycloalkyl” and the like. refers to a heterocycloalkyl group derived from a 3- to 10-membered heterocycloalkane moiety containing a heteroatom by extraction of one hydrogen atom. Examples include, but are not limited to, aziridinyl, azetidinyl, azacyclofentanyl, azacyclohexanyl, azepanyl, azocanyl, azoninyl, and the like.

The term “oxo” as used herein refers to a substituent in which an oxygen atom is bonded to a carbon atom or a nitrogen atom. Carbonyl is mentioned as a specific example of the structure in which the oxygen atom couple|bonded with the carbon atom, and N-oxide is mentioned as a specific example of the group which the oxygen atom couple|bonded with the nitrogen atom.

"Halo" refers to fluoro, chloro, bromo and iodo. “C _1-6 alkoxy” refers to (C _{1-6 alkyl} )O—, wherein C _{1-6 alkyl} is as defined herein. “Halo _- C _1-6 alkyl” refers to halo-(C _{1-6 alkyl} )-, wherein C _1-6 alkyl is as defined herein. Halo-C _1-6 alkyl is a perhalogenated C _1-6 alkyl in which all hydrogen atoms in the C _1-6 alkyl are replaced by halogen, such as —CF ₃ , —CH ₂ CF ₃ , —CF ₂ CF ₃ , —CH ₂ CH ₂ CF ₃ , and the like.

The term "5- to 8-membered lactam group" as described herein includes, for example, a "5- to 7-membered lactam group" including -C(=O) in the ring by extraction of one hydrogen atom. -N refers to a group derived from a lactam containing a group. Examples include, but are not limited to:

The compounds of the present invention may be prepared or used as pharmaceutically acceptable salts thereof. This can be done by any means of salt formation known in the art. For example, the pharmaceutically acceptable salt can be an acid addition salt, such as an inorganic acid addition salt or an organic acid addition salt. For example, the pharmaceutically acceptable salt may be a salt formed by substituting a metal ion for an acidic proton in the compound, or a salt formed by coordinating the compound with an organic base or an inorganic base.

The term “pharmaceutically acceptable” as used herein in reference to a formulation, composition or ingredient does not have a lasting detrimental effect on the general health of the subject being treated or abrogate the biological activity or properties of the compound, and This means that it is relatively non-toxic.

The term “effective amount” or “therapeutically effective amount” as used herein refers to a sufficient amount of an agent or compound being administered that will alleviate to some extent one or more symptoms of the disease or condition being treated. The result may be reduction and/or alleviation of the signs, symptoms or causes of a disease, or any other desirable alteration of a biological system. For example, an “effective amount” for therapeutic use is an amount necessary to provide a clinically significant reduction in disease symptoms without undue adverse side effects. An appropriate "effective" amount in an individual case can be determined using techniques such as dose escalation studies. The term “therapeutically effective amount” includes, for example, a prophylactically effective amount. An “effective amount” of a compound disclosed herein is an amount effective to achieve the desired pharmacological effect or therapeutic improvement without undue adverse side effects. It is understood that an "effective amount" or "therapeutically effective amount" can vary from subject to subject, due to variations in the metabolism of the compound, the age, weight, general condition, condition being treated, the severity of the condition being treated, and the judgment of the prescribing physician. do. By way of example only, a therapeutically effective amount can be determined by routine experimentation, including, but not limited to, dose escalation clinical trials.

As used herein, the terms “inhibit”, “inhibiting” and “inhibitor” for a kinase refer to inhibition of the activity of a CDK4/6 kinase.

For general methods for preparing compounds as disclosed herein, reference may be made to reactions known in the art. One of ordinary skill in the art can reasonably select appropriate reagents and conditions to adapt or modify reactions known in the art for introducing individual moieties into the structure of a compound as disclosed herein.

Starting materials used in the synthesis of the compounds described herein can be synthesized or obtained from Aldrich Chemical Co., Ltd. (Milwaukee, Wisconsin), Bachem (Torrance, California), or Sigma Chemical Co. (St. Louis, Mo.) from commercial sources such as, but not limited to. The compounds described herein, and other related compounds having different substituents, are described, for example, in March, ADVANCED ORGANIC CHEMISTRY, 4th Ed., (Wiley 1992); Carey and Sundberg, ADVANCED ORGANIC CHEMISTRY, 4th Ed., Vols. A and B (Plenum 2000, 2001); Green and Wuts, PROTECTIVE GROUPS IN ORGANIC SYNTHESIS, 3rd Ed., (Wiley 1999); Fieser's Reagents for Organic Synthesis, Volumes 1-17 (John Wiley and Sons, 1991); Rodd's Chemistry of Carbon Compounds, Volumes 1-5 and Supplementals (Elsevier Science Publishers, 1989); Organic Reactions, Volumes 1-40 (John Wiley and Sons, 1991); and Larock's Comprehensive Organic Transformations (VCH Publishers Inc., 1989)), which is incorporated herein by reference in its entirety.

The product of the reaction can be isolated and purified, if desired, using conventional techniques including, but not limited to, filtration, distillation, crystallization, chromatography, and the like. These products can be characterized using conventional means, including physical constants and spectral data.

The compounds described herein can be prepared as a single isomer or a mixture of isomers by the synthetic methods described herein.

The compounds described herein may have one or more stereocenters and each center exists in either the R or S configuration. The compounds presented herein include all diastereomeric, enantiomeric and epimeric forms, as well as suitable mixtures thereof. Stereoisomers can, if desired, be obtained by methods known in the art, for example by separation of stereoisomers by chiral chromatography columns.

Diastereomeric mixtures can be separated into individual diastereomers on the basis of differences in physicochemical properties by known methods such as, for example, chromatography and/or fractional crystallization. In one embodiment, enantiomers can be separated by a chiral chromatography column. In some other embodiments, enantiomers convert an enantiomer mixture to a diastereomeric mixture by reaction with an appropriate optically active compound (eg, an alcohol), separate the diastereomers, and convert the individual diastereomers to the corresponding can be separated by conversion (eg, hydrolysis) to the pure enantiomer. All such isomers, including diastereomers, enantiomers, and mixtures thereof, are considered components of the compositions described herein.

The methods and formulations described herein include the use of N-oxides, crystalline forms (also known as polymorphs), or pharmaceutically acceptable salts of the compounds described herein, as well as active metabolites of these compounds having the same type of activity. includes In some circumstances, compounds may exist as tautomers. All tautomers are included within the scope of the compounds presented herein. In addition, the compounds described herein may exist in unsolvated as well as solvated forms using pharmaceutically acceptable solvents such as water, ethanol, and the like. Solvated forms of the compounds provided herein are also considered to be disclosed herein.

The unoxidized form forms sulfur, sulfur dioxide, triphenyl phosphine, lithium borohydride, sodium borohydride at 0-80° C. in a suitable inert organic solvent such as, but not limited to, acetonitrile, ethanol, aqueous dioxane, and the like. , phosphorus trichloride, tribromide, and the like can be prepared from N-oxides by treatment with a reducing agent such as, but not limited to.

In some embodiments, a compound described herein is prepared as a prodrug. "Prodrug" refers to an agent that is converted in vivo to the parent drug. Prodrugs are often useful because they are easier to administer than the parent drug in some situations. For example, in some cases the active compound itself is not bioavailable by oral administration, and prodrugs may be used to achieve this goal. The prodrug may have improved solubility in the pharmaceutical composition compared to the parent drug. Prodrugs can be designed as reversible drug derivatives to enhance drug transport into site-specific tissues. In some embodiments, the design of the prodrug increases effective aqueous solubility. See, eg, Fedorak, et al., Am. J. Physiol, 269: G210-218 (1995); McLoed, et al., Gastroenterol, 106: 405-413 (1994); Hochhaus, et al., Biomed. Chrom., 6:283-286 (1992); J. Larsen and H. Bundgaard, Int. J. Pharmaceutics, 37, 87 (1987); J. Larsen, et al., Int. J. Pharmaceutics, 47, 103 (1988); Sinkula, et al., J. Pharm. Sd., 64:181-210 (1975); T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, the A.C.S. Symposium Series, Vol. 14; and Edward B. Roche, Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987, all of which are incorporated herein by reference in their entirety. Non-limiting examples of prodrugs include administration as esters (“prodrugs”) to facilitate delivery across cell membranes, where water solubility is detrimental to mobility, and then once metabolized inside cells where water solubility is beneficial to the active substance, a carboxylic acid. A compound as described herein that is hydrolyzed. A further example of a prodrug may be a short peptide (polyamino acid) conjugated to an acid group from which the peptide is metabolized to reveal an active moiety. In certain embodiments, upon administration in vivo, a prodrug is chemically converted into a biologically, pharmaceutically or therapeutically active form of the compound. In certain embodiments, a prodrug is enzymatically metabolized by one or more steps or processes to a biologically, pharmaceutically or therapeutically active form of the compound. A pharmaceutically active compound can be modified such that, upon in vivo administration, the active compound is regenerated to produce a prodrug. Prodrugs can be designed to alter the metabolic stability or transport properties of the drug, hide side effects or toxicity, improve the effectiveness of the drug, or alter other properties or properties of the drug. Based on knowledge of in vivo pharmacodynamic processes and drug metabolism, once a pharmaceutically active compound is known, one of skill in the art can design prodrugs of the compound (see, e.g., Nogrady (1985) Medicinal Chemistry A Biochemical Approach, Oxford University Press, New York, pages 388-392; Silverman (1992), The Organic Chemistry of Drug Design and Drug Action, Academic Press, Inc., San Diego, pages 352-401; and Saulnier, et al., (1994) ), Bioorganic and Medicinal Chemistry Letters, Vol. 4, p. 1985].

Prodrug forms of the compounds described herein that are metabolized in vivo to yield the derivatives described herein are included within the scope of the claims. In some cases, some of the compounds described herein may be prodrugs or other derivatives of the active compounds.

The compounds described herein are described herein in their molecular formulas and structural formulas, except for the fact that one or more atoms are replaced by a nuclide belonging to the same element but having an atomic mass or mass number different from the atomic mass or mass number ordinarily found in nature. compounds comprising the same isotopes as recited are encompassed. For example, when hydrogen is present at any position in the compounds described herein, it includes instances where an isotope of hydrogen (eg, protium, deuterium, tritium, etc.) occurs at that position. Examples of isotopes that can be incorporated into the compounds described herein include, for example, ² H, ³ H, ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁸ O, ¹⁷ O, ³⁵ S, ¹⁸ F, ³⁶ Cl, each isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine and chlorine such as, but not limited to. Compounds described herein containing certain isotopes (eg, radioactive isotopes such as ³ H and ¹⁴ C) are useful in drug and/or substrate tissue distribution assays.

In further or additional embodiments, the compounds described herein are metabolized upon administration to an organism in need thereof to produce a metabolite and used to produce a desired effect, including a desired therapeutic effect.

The compounds described herein can be formed and/or used as pharmaceutically acceptable salts. Types of pharmaceutically acceptable salts include, but are not limited to: (1) combining the compound in its free base form with a pharmaceutically acceptable inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, metaphosphoric acid, etc. react; or organic acids such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, adipic acid, sebacic acid, succinic acid, malic acid, maleic acid, fumaric acid, trifluoroacetic acid, tartaric acid, Citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, toluene Sulfonic acid, 2-naphthalenesulfonic acid, 4-methylbicyclo-[2.2.2]oct-2-ene-1-carboxylic acid, glucoheptonic acid, hippuric acid, gentisic acid, 4,4'-methylenebis-( 3-hydroxy-2-ene-1-carboxylic acid), nicotinic acid, 3-phenylpropionic acid, trimethylacetic acid, tert-butylacetic acid, lauryl sulfate, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid, acid addition salts formed by reacting with the like; (2) an acidic proton present in the parent compound is a metal ion, such as an alkali metal ion (eg, lithium, sodium, or potassium), an alkaline earth ion (eg, magnesium or calcium), or an aluminum ion salts formed when replaced with; or (3) a salt formed by coordination with an organic or inorganic base. Acceptable organic bases include ethanolamine, diethanolamine, triethanolamine, trimethylamine, N-methylglucamine, and the like. Acceptable inorganic bases include aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate, sodium hydroxide, and the like.

Corresponding counterions of pharmaceutically acceptable salts include, but are not limited to, ion exchange chromatography, ion chromatography, capillary electrophoresis, inductively coupled plasma, atomic absorption spectroscopy, mass spectrometry, or any combination thereof. It can be analyzed and identified using a variety of methods that do not

The salt may be recovered using one or more of the following techniques: filtration, precipitation with a nonsolvent, followed by filtration, evaporation of the solvent, or lyophilization in the case of an aqueous solution.

It should be understood that reference to a pharmaceutically acceptable salt includes solvent addition forms or crystalline forms thereof, in particular solvates or polymorphs. Solvates contain either stoichiometric or non-stoichiometric amounts of solvent and may be formed during the crystallization process with pharmaceutically acceptable solvents such as water, ethanol, and the like. Hydrates are formed when the solvent is water, and alcoholates are formed when the solvent is alcohol. Solvates of the compounds described herein can be conveniently prepared or formed during the processes described herein. In addition, the compounds provided herein can exist in unsolvated and solvated forms. In general, solvated forms are considered equivalent to unsolvated forms for the purposes of the compounds and methods provided herein.

The compounds described herein can be in a variety of forms, including, but not limited to, amorphous forms, spherical forms, and nano-particulate forms. The compounds described herein also include crystalline forms, also known as polymorphs. Polymorphs contain different crystal packing arrangements of the same elemental composition of a compound. Polymorphs generally differ in X-ray diffraction pattern, infrared spectrum, melting point, density, hardness, crystal shape, optical and electrical properties, stability and solubility. The monocrystalline form may predominate due to various factors such as recrystallization solvent, crystallization rate and storage temperature.

Screening and characterization of pharmaceutically acceptable salts, polymorphs and/or solvates can be accomplished using a variety of techniques including, but not limited to, thermal analysis, x-ray diffraction, spectroscopy, vapor adsorption, and microscopy. there is. Thermal analysis methods address thermochemical degradation or thermophysical processes including, but not limited to, polymorphic transitions and these methods can be used to analyze relationships between polymorphic forms, to determine weight loss, to find glass transition temperatures, or to use excipients It can be used for sex studies. Such methods include, but are not limited to, differential scanning calorimetry (DSC), modulated differential scanning calorimetry (MDCS), thermogravimetric analysis (TGA), and thermogravimetry and infrared analysis (TG/IR). X-ray diffraction methods include, but are not limited to, single crystal and powder diffractometers and synchrotron sources. Various spectroscopic techniques used include, but are not limited to, Raman, FTIR, UVIS, and NMR (liquid and solid state). Various microscopy techniques include polarized light microscopy, scanning electron microscopy (SEM) using energy dispersive X-ray analysis (EDX), environmental scanning electron microscopy using EDX (in a gas or water vapor atmosphere), IR microscopy , and Raman microscopy.

Carriers herein include conventional diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption enhancers, surfactants, adsorption carriers, lubricants, and, if necessary, flavoring agents, sweetening agents, and the like, commonly used in the field of pharmacy. includes The pharmaceutical of the present invention can be prepared in various forms such as tablets, powders, granules, capsules, oral liquids, injections, and the like. The medicaments in various dosage forms can be prepared by methods conventional in the field of pharmaceuticals.

As used herein, IC ₅₀ is an amount, concentration or dose of a particular test compound that achieves 50% inhibition of a maximal response in an assay that measures response, eg, inhibition of a CDK kinase such as CDK4/6 kinase. refers to

Example

The following specific and non-limiting examples are to be construed as illustrative only and do not limit the present disclosure in any way. Without further explanation, it is believed that those skilled in the art will be able to make the most of the present invention based on the description herein.

synthesis of compounds

The following abbreviations are used in the following synthetic schemes.

Boc: t-butoxycarbonyl;

Et: ethyl;

H: time;

rt/RT: room temperature;

Me: methyl;

MeOH: methanol;

DIPEA: N,N-diisopropylethylamine;

DCM: dichloromethane;

NMP: N-methylpyrrolidone;

TEA: triethylamine;

TFA: trifluoroacetic acid;

THF: tetrahydrofuran;

HATU: 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethyluronium hexafluorophosphate;

DMF: dimethylformamide;

NBS: N-bromosuccinimide;

NIS: N-iodosuccinimide;

DPPF PdCl ₂ : [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride;

XantPhos: 4,5-bisdiphenylphosphino-9,9-dimethylxanthene;

Pd ₂ (dba) ₃ : tris(dibenzylideneacetone)dipalladium.

Synthesis Scheme I

Step 1:

To a 1 L two-necked flask were added 100 g of compound 1-1 and 91 g of compound 1-2 , followed by 500 mL of dioxane as solvent. A water separator and condenser were connected. 9 mL of hydrochloric acid solution (2 mol/L) was added. The mixture was heated to 115° C. and reacted for 1 hour. Then 2 mL of HCl solution (2 mol/L) was added and the reaction was carried out at a controlled temperature of 95° C. overnight. After the reaction was complete, the reaction mixture was cooled to room temperature. The pH was adjusted to an alkaline value by addition of saturated sodium bicarbonate solution. The mixture was concentrated to remove dioxane and extracted with ethyl acetate. The resulting organic layer was dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography (gradient elution, acetone: n-hexane = 1:8 to 1:5) to give 40 g of product.

Step 2:

To a two-neck flask was added 40 g of compound 1-3 , followed by 250 mL of acetonitrile for (ultrasound) dissolution. Under the protection of argon, 20 mL of compound 1-4 was slowly added dropwise in an ice - salt bath, and the reaction was carried out for 1 hour. After the reaction was completed, 40 mL of DMF was added and the reaction was carried out for 1 hour with stirring in an ice-salt bath. After the reaction was completed, the reaction mixture was poured into ice water and extracted with ethyl acetate. The resulting organic layer was dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography (gradient elution, ethyl acetate: n-hexane = 1:10 to 1:5) to give 38 g of product.

Step 3:

To a 1 L flask were added 38 g of compound 1-5 , followed by 665 mL of acetonitrile as solvent. The temperature was reduced to -30°C under the protection of argon. Then, 27.5 g of compound 1-6 was slowly added and reacted for 5 minutes with stirring. The reaction was allowed to warm to room temperature for 2 hours. After the reaction was complete, ethyl acetate and saturated brine were added for extraction. The resulting organic layer was dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography (gradient elution, ethyl acetate: n-hexane = 1:10 to 1:5) to give 47 g of product.

Step 4:

To a 2 L flask was added 47 g of compound 1-7 , followed by 940 mL of ethanol as a solvent. 470 mL of aqueous ammonia was added, and 83.5 mL of aqueous hydrogen peroxide (30%) was slowly added dropwise at room temperature. After addition, the mixture was stirred overnight. After the reaction was complete, the mixture was concentrated to remove the ethanol and filtered to give a light yellow solid. The solid was washed with pure water, then methyl tert-butyl ether and n-hexane (1:1) to give a white solid, which was dried in vacuo to give 37 g of product.

Step 5:

To a 100 mL flask, 6 g of compound 1-8 , and then 10 mL of dichloromethane were added for dissolution. Then, 20 mL of trifluoroacetic acid was added dropwise and reacted for 2 hours with stirring at room temperature. After the reaction was complete, the mixture was concentrated to remove the solvent and excess trifluoroacetic acid, adjusted to pH 7-8 with saturated sodium bicarbonate, and extracted with ethyl acetate. The resulting organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give 3.6 g of product.

Step 6:

To a 250 mL flask were added 3 g of compound 1-9 , 100 mL of dichloromethane, and then 2.4 g of compound 1-10 . After stirring at room temperature, triethylamine (4.5 g) was slowly added dropwise. Upon completion of the reaction by monitoring by TLC, the reaction mixture was concentrated and ready for direct use.

Step 7:

To a 250 mL flask was added crude compound 1-11 obtained from step 6. Then, 100 mL of ethanol was added as a solvent, and 1.7 g of potassium hydroxide was added. The mixture was heated to reflux overnight. After the reaction was completed, acetic acid was added to adjust the pH to 5-6. A solid precipitated and was filtered. The filter cake was washed with water followed by a small amount of diethyl ether and dried under infrared light to give 2.5 g of product.

Step 8:

To a 100 mL pressure-resistant tube was added 2.5 g of compound 1-13 , followed by 20 mL of compound 1-14 and 10 mL of DIPEA. The mixture was heated to 125° C. and reacted for 48 hours. After the reaction was completed, the reaction mixture was concentrated to remove excess phosphorus oxychloride and DIPEA. 100 mL of ethyl acetate was added for dissolution and dilution. A small number of ice cubes were added in an ice-water bath, then the pH was adjusted to 7-8 by slowly dropwise addition of saturated sodium bicarbonate solution. Saturated sodium chloride solution was added and the mixture was extracted with ethyl acetate. The resulting organic layer was dried, filtered, concentrated, and purified by column chromatography to obtain 1.5 g of product.

Step 9:

To a 100 mL flask were added 3.9 g of compound 1-15 and 0.9 g of sodium borohydride, followed by 40 mL of THF as solvent. After addition of 2 mL of isopropanol, the reaction was carried out at room temperature with stirring for 1 hour. After the reaction was complete, the reaction mixture was filtered to remove solids, and washed with DCM. The filtrate was concentrated to remove excess THF, isopropanol, and DCM. Then, DCM and 5 g of DDQ as solvents were added, and the reaction was conducted at room temperature for 30 minutes. After the reaction was completed, the reaction mixture was filtered to remove solids. Saturated sodium bicarbonate solution was added to the filtrate, which was extracted with DCM, dried, filtered, concentrated and purified by column chromatography to give 3 g of product.

Step 10:

In a 50 mL flask, 1 g of compound 1-16 , 0.9 g of compound 1-17 , 0.3 g of DPPF PdCl ₂ , and 0.9 g of potassium carbonate, followed by 10 mL of toluene and 1 mL of water as a solvent added. The reaction was carried out overnight at 100° C. under the protection of argon. After the reaction was completed, ethyl acetate and saturated sodium chloride solution were added for extraction. The resulting organic layer was dried, filtered, concentrated, and purified by column chromatography to give 430 mg of product.

Step 11:

In a 100 mL flask, 430 mg of compound 1-18 , 565 mg of compound 1-19 , 106 mg of XantPhos, 84 mg of Pd ₂ (dba) ₃ , and 900 mg of sodium tert-butoxide, and then 20 mL of toluene was added as solvent. The mixture was heated to 105° C. and reacted overnight under the protection of argon. After the reaction was completed, saturated sodium chloride solution was added, and the mixture was extracted with ethyl acetate. The resulting organic layer was dried, filtered, concentrated and purified by column chromatography to give 200 mg of product.

Step 12:

To a 50 mL flask were added 150 mg of compound 1-20 , followed by 15 mL of tetrahydrofuran as a solvent. After stirring at 0° C. for 10 minutes, 56 mg of NBS was slowly added thereto and reacted for 15 minutes with stirring at 0° C. After monitoring the completion of the reaction, saturated sodium chloride solution was added and the mixture was extracted with ethyl acetate. The resulting organic layer was dried, filtered, concentrated and purified by column chromatography to give 160 mg of product.

Step 13:

To a 50 mL flask were added 100 mg of compound 1-22 , 42 mg of compound 1-23 , 13 mg of DPPF PdCl ₂ , and 38 mg of potassium carbonate. Then, 15 mL of dioxane was added as a solvent and 1.5 mL of water was added. The mixture was heated to 110° C. and reacted overnight under the protection of argon. After the reaction was completed, saturated sodium chloride solution was added, and the mixture was extracted with ethyl acetate. The resulting organic layer was dried, filtered, concentrated and purified by thin layer chromatography on a thick preparative plate to give 10 mg of product.

Step 14:

10 mg of compound 1-24 in a 25 mL flask. 5 mL of dichloromethane was added for dissolution, followed by 0.5 mL of HCl/MeOH solution (3 N). The mixture was stirred and reacted at room temperature for 2 hours. After the reaction was complete, the reaction mixture was concentrated to remove the solvent and excess HCl/MeOH and dried in vacuo to give 9 mg of product.

Synthesis Scheme II

Step 1:

To a 50 mL pressure-resistant tube was added 2.2 g of compound 1-16 , 10 mL of tetrahydrofuran was added as a solvent, and then 5 mL of aqueous ammonia (mass fraction: 28%) was added. The mixture was heated to 100° C. and reacted overnight. After completion of the reaction, the reaction mixture was directly concentrated to remove the solvent and excess aqueous ammonia to obtain 2.3 g of a product.

Step 2:

To a 50 mL flask were added 2.3 g of compound 2-2 and 15 mL of compound 2-3 . The mixture was heated to 100° C. and reacted for 4 hours. After the reaction was completed, saturated sodium chloride solution was added, and the mixture was extracted with ethyl acetate. The resulting organic layer was dried, filtered and concentrated to give the crude product, which was washed with a mixture of methyl tert-butyl ether and n-hexane (1:1) and dried in vacuo to give 2.2 g of product .

Step 3:

To a 10 mL flask were added 200 mg of compound 2-4 , 211 mg of compound 2-5 , 40 mg of catalyst (CAS: 1447963-75-8), and 2.5 mL of potassium acetate solution (0.5 N). The mixture was heated to 100° C. and reacted under the protection of argon for 2 hours. After the reaction was completed, saturated sodium chloride solution was added, and the mixture was extracted with ethyl acetate. The resulting organic layer was dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography to give 110 mg of product.

Step 4:

To a 50 mL flask were added 110 mg of compound 2-6 and 20 mL of tetrahydrofuran. The mixture was stirred in an ice-water bath for 10 minutes. Then, 86 mg of compound 21 was added slowly. After monitoring the completion of the reaction, saturated sodium chloride solution was added and the mixture was extracted with ethyl acetate. The resulting organic layer was dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography to give 150 mg of product.

Step 5:

To a 50 mL flask were added 150 mg of compound 2-7 , 149 mg of compound 2-8 , 37.4 mg of DPPF PdCl ₂ , and 106 mg of potassium carbonate. Then, 15 mL of dioxane was added as a solvent and 1.5 mL of water was added. The mixture was heated to 110° C. and reacted overnight under the protection of argon. After the reaction was completed, saturated sodium chloride solution was added, and the mixture was extracted with ethyl acetate. The resulting organic layer was dried, filtered, concentrated and purified by column chromatography to give 80 mg of product.

Step 6:

To a 50 mL flask was added 80 mg of compound 2-9 . 10 mL of tetrahydrofuran was added as a solvent, followed by the addition of 5 mL of sodium hydroxide solution (2 N). The mixture was heated to reflux and reacted for 4 hours. After the reaction was completed, saturated sodium chloride solution was added, and the mixture was extracted with ethyl acetate. The resulting organic layer was dried over anhydrous sodium sulfate, filtered, concentrated and dried in vacuo to give 50 mg of product.

Step 7:

In a 50 mL flask, 50 mg of compound 2-11 , 129 mg of compound 2-12 , 12.8 mg of XantPhos, 10 mg of Pd ₂ (dba) ₃ , and 32 mg of sodium tert-butoxide, and then 15 mL of toluene was added as solvent. The mixture was heated to 110° C. and reacted overnight under the protection of argon. After the reaction was completed, saturated sodium chloride solution was added, and the mixture was extracted with ethyl acetate. The resulting organic layer was dried over anhydrous sodium sulfate, filtered, concentrated and purified by thin layer chromatography on a thick preparative plate to give 10 mg of product.

Step 8:

To a 25 mL flask was added 10 mg of compound 2-13 . 5 mL of dichloromethane was added for dissolution, followed by 0.5 mL of HCl/MeOH solution (3 N). The mixture was stirred and reacted at room temperature for 2 hours. After the reaction was complete, the reaction mixture was concentrated to remove the solvent and excess HCl/MeOH and dried in vacuo to give 9 mg of product.

Synthesis Scheme III

Step 1:

In a 100 mL flask, 500 mg of compound 2-2 , 964 mg of compound 3-1 , 135.8 mg of XantPhos, 107.5 mg of Pd ₂ (dba) ₃ , and 338 mg of sodium tert-butoxide, and then 40 mL of toluene was added as solvent. The mixture was heated to 105° C. and reacted overnight under the protection of argon. After the reaction was completed, saturated sodium chloride solution was added, and the mixture was extracted with ethyl acetate. The resulting organic layer was dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography to give 480 mg of product.

Step 2:

To a 50 mL flask were added 230 mg of compound 3-2 , 121 mg of compound 1-17 , 35 mg of DPPF PdCl ₂ , and 101 mg of potassium carbonate. 15 mL of dioxane was added as solvent, followed by 1.5 mL of water. The mixture was heated to 110° C. and reacted overnight under the protection of argon. After the reaction was completed, saturated sodium chloride solution was added, and the mixture was extracted with ethyl acetate. The resulting organic layer was dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography to give 150 mg of product.

For other steps, see steps 12 , 13 , and 14 of Synthesis Scheme I.

Synthesis Scheme IV

Step 1:

To a 100 mL flask was added 1 g of compound 2-2 , followed by 50 mL of DMF for dissolution. The mixture was stirred in an ice-water bath for 10 minutes, then 1.06 g of compound 4-1 were added (in three portions, 15 minutes apart). After 0.5 h reaction, the reaction was monitored. After the reaction was completed, saturated sodium chloride solution was added, and the mixture was extracted with ethyl acetate. The resulting organic layer was dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography to give 0.9 g of product.

Step 2:

To a 100 mL flask were added 0.9 g of compound 4-2 , 610 mg of compound 2-8 , 194 mg of DPPF PdCl ₂ , and 732 mg of potassium carbonate. 50 mL of dioxane was added as a solvent, followed by 5 mL of water. The mixture was heated to 72° C. and reacted overnight under the protection of argon. After the reaction was completed, saturated sodium chloride solution was added, and the mixture was extracted with ethyl acetate. The resulting organic layer was dried, filtered, concentrated, and purified by column chromatography to give 380 mg of product.

Step 3:

In a 100 mL flask, 380 mg of compound 4-3 , 559 mg of compound 3-1 , 78 mg of XantPhos, 62 mg of Pd ₂ (dba) ₃ , and 131 mg of sodium tert-butoxide, and then 30 mL of toluene was added as solvent. The mixture was heated to 105° C. and reacted overnight under the protection of argon. After the reaction was completed, saturated sodium chloride solution was added, and the mixture was extracted with ethyl acetate. The resulting organic layer was dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography to give 390 mg of product.

Step 4:

To a 100 mL flask were added 43 mg of compound 4-4 , 52 mg of compound 4-5 , and 1 mg of DMAP, followed by acetonitrile as solvent. The reaction was carried out at room temperature overnight. After the reaction was completed, saturated sodium chloride solution was added, and the mixture was extracted with ethyl acetate. The resulting organic layer was dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography to give 35 mg of product.

Step 5:

To a 100 mL flask were added 35 mg of compound 4-6 , 21 mg of compound 4-7 , 4 mg of DPPF PdCl ₂ , and 9.5 mg of potassium fluoride, followed by 5 mL of DMSO as solvent. The mixture was heated to 130° C. in the microwave and reacted under the protection of argon for 1 hour. After the reaction was completed, saturated sodium chloride solution was added, and the mixture was extracted with ethyl acetate. The resulting organic layer was dried over anhydrous sodium sulfate, filtered, concentrated and purified by thin layer chromatography to give 7 mg of product.

Step 6:

To a 25 mL flask was added 7 mg of compound 4-8 . 5 mL of dichloromethane was added for dissolution, followed by the addition of 0.5 mL of TFA. The reaction was carried out at room temperature with stirring for 2 hours. After the reaction was complete, the reaction mixture was concentrated to remove the solvent and excess TFA and dried in vacuo to give 7 mg of product.

Synthesis Scheme V

Step 1:

To a microwave tube was added compound 4-4 (1.0 g), bis(pinacholate)diboron (1.19 g), PdCl ₂ (dppf) (57.11 mg), potassium acetate (459.63 mg), and DMF (15 mL). . The reaction was carried out under the protection of argon in a microwave at 135° C. for 1.5 hours. After the reaction was complete, EA and H ₂ O were added to extract, and the layers were separated. The organic phase was dried to remove water, spin-dried and purified via column (gradient elution, eluent: EA:Hex = 1:5 to 1:1) to give compound 5-2 (787.6 mg).

Step 2:

Compound 5-2 (50 mg), compound 5-3 (34.18 mg), PdCl ₂ (dppf) (5.32 mg), potassium fluoride (21.12 mg), DMSO (5 mL), and water (0.5 mL) in a microwave tube was added. The reaction was carried out under the protection of argon in a microwave at 130° C. for 1.5 hours. After the reaction was complete, EA and H ₂ O were added to extract, and the layers were separated. The organic phase was rotary dried and purified by thin layer chromatography to give compound 5-4 (7 mg).

Step 3:

7 mg of compound 5-4 was added to a 25 mL flask. 5 mL of dichloromethane was added for dissolution, followed by the addition of 0.5 mL of TFA. The mixture was stirred and reacted at room temperature for 2 hours. After the reaction was complete, the reaction mixture was concentrated to remove the solvent and excess TFA and dried in vacuo to give 6.3 mg of product.

Synthesis Scheme VI

Step 1:

In a 100 mL reaction flask, compound 1-16 (500 mg), dimethylamine hydrochloride (210.47 mg), Pd ₂ (dba) ₃ (147.72 mg), Xantphos (186.68 mg), K ₃ PO ₄ (1.0 g), and toluene (15 mL) were added. CO gas was charged, and the mixture was heated to 105° C. and stirred overnight. The reaction was filtered with suction through a pad of celite and washed with DCM. The filtrate was rotary dried and purified via column (gradient elution, eluent: EA:Hex = 1:3-3:1) to give product 6-2 (250 mg).

Step 2:

To the reaction flask was added compound 6-2 (250 mg), THF (9 mL), and MeOH (3 mL) in an ice-water bath, and NBS (200 mg) was added in portions. The mixture was stirred for 1 hour. After removing the ice bath and raising the temperature to room temperature, the mixture was stirred for an additional hour. After the reaction was complete, EA and H ₂ O were added, secured, and the layers were separated. The organic phase was dried, spin dried and purified via column (gradient elution, eluent: EA:Hex = 1:1-3:1) to give compound 6-3 (125 mg).

Step 3:

Compound 6-3 (150 mg), 1-cyclopentene borate pinacol (143.86 mg), catalyst PdCl ₂ (dppf) (36.16 mg), and potassium carbonate (170.74 mg), followed by dioxane (4) in a microwave tube mL) and water (0.4 mL) were added as solvents. The reaction was carried out under the protection of argon in a microwave at 110° C. for 1 hour. After the reaction was complete, EA and H ₂ O were added, secured, and the layers were separated. The organic phase was dried over anhydrous sodium sulfate, spin dried and purified via column (gradient elution, eluent: EA:Hex = 1:1-3:1) to give compound 6-4 (100 mg).

Step 4:

450 mg of compound 6-4 was added to a 20 mL pressure-resistant tube. Tetrahydrofuran (4 mL) was added as a solvent, followed by 8 mL of aqueous ammonia (mass fraction: 28%). The mixture was heated to 100° C. and reacted overnight. After completion of the reaction, the reaction mixture was directly concentrated to remove the solvent and excess aqueous ammonia to obtain 300 mg of product.

Step 5:

In a reaction flask, compound 6-5 (40 mg), compound 6-6 (67.63 mg), Pd ₂ (dba) ₃ (6.75 mg), Xantphos (9.38 mg), sodium tert-butoxide (35.42 mg), and toluene (10 mL) was added. The mixture was heated to 105° C. and stirred overnight under the protection of argon. After the reaction was completed, the solid was filtered off, and the filtrate was rotary dried and purified by thin layer chromatography to obtain compound 6-7 (43 mg).

Step 6:

To a 25 mL flask was added 43 mg of compound 6-7 . 5 mL of dichloromethane was added for dissolution, followed by the addition of 1 mL of TFA. The mixture was stirred and reacted at room temperature for 2 hours. After the reaction was complete, the reaction mixture was concentrated to remove the solvent and excess TFA and dried in vacuo to give 60 mg of product.

Synthesis Scheme VII

Step 1:

In a 50 mL flask 150 mg of compound 4-3 , 168 mg of compound 1-17 , 39 mg of DPPF PdCl ₂ , and 148 mg of potassium carbonate, followed by 10 mL of toluene and 1 mL of water as solvent added. The reaction was carried out at 110° C. under the protection of argon overnight. After the reaction was completed, ethyl acetate and saturated sodium chloride solution were added for extraction. The resulting organic layer was dried, filtered, concentrated and purified by column chromatography to give 90 mg of product.

Step 2:

To a 100 mL reaction flask were added 90 mg of compound 7-1 , an appropriate amount of Pd/C catalyst, followed by 8 mL of tetrahydrofuran and 15 mL of methanol. The mixture was purged with hydrogen and reacted for 2 hours at room temperature under a hydrogen atmosphere. After the reaction was complete, the reaction mixture was filtered, the filtrate was concentrated and dried in vacuo to give an intermediate product.

The obtained intermediate was dissolved in 15 mL of dichloromethane, then 100 mg of DDQ was added. The mixture was stirred and reacted at room temperature for 1 hour. After the reaction was completed, the reaction mixture was filtered. Saturated sodium bicarbonate solution was added to the filtrate, and this was extracted with ethyl acetate to obtain an organic layer. The organic layer was dried, concentrated and purified by column chromatography to give 50 mg of product ( 7-2 ).

Step 3:

To the reaction flask were added 20 mg of compound 7-2 , 37 mg of compound 3-1 , 4 mg of Pd ₂ (dba) ₃ , 5 mg of Xantphos, 14 mg of sodium tert-butoxide, and 10 mL of toluene. did. Under the protection of argon, the mixture was heated to 105° C. and stirred overnight. After the reaction was completed, the solid was filtered off, and the filtrate was rotary dried and purified by thin layer chromatography to obtain 7 mg of compound 7-3 .

Step 4:

7 mg of compound 7-3 was added to a 25 mL flask. 5 mL of dichloromethane was added for dissolution, followed by the addition of 1 mL of TFA. The mixture was stirred and reacted at room temperature for 2 hours. After the reaction was complete, the mixture was concentrated to remove the solvent and excess TFA and dried in vacuo to give 8.2 mg of product 7-4 .

The salt-forming compounds obtained in the above synthetic schemes I to VII can be prepared in non-salt-forming forms by methods known in the art using, for example, the following desalting schemes I or II.

Desalination Scheme I

To the flask was added the salt form of compound I. Methanol was added as solvent, followed by addition of an excess of saturated sodium bicarbonate solution. The mixture was stirred and reacted at room temperature for 4 hours. After the reaction was completed, the reaction mixture was concentrated to remove a portion of methanol and extracted with dichloromethane. The resulting organic layer was dried over anhydrous sodium sulfate, filtered, concentrated to remove the solvent, and dried in vacuo to give the target compound I.

A more specific example (compound of Example 111 as described below) is as follows:

13 mg of compound 8-1 was added to a 25 mL flask. 3 mL of methanol was added as solvent, followed by 2 mL of saturated sodium bicarbonate solution. The mixture was stirred and reacted at room temperature for 4 hours. After the reaction was completed, the reaction mixture was concentrated to remove a portion of methanol and extracted with dichloromethane. The resulting organic layer was dried over anhydrous sodium sulfate, filtered, concentrated to remove the solvent, and dried in vacuo to give 9.4 mg of the target compound 8-2 .

Desalination Scheme II

The salt form of compound II was added to the flask. Methanol was added as solvent, followed by addition of an excess of saturated sodium bicarbonate solution. The mixture was stirred and reacted at room temperature for 4 hours. After the reaction was completed, the reaction mixture was concentrated to remove a portion of methanol and extracted with dichloromethane. The resulting organic layer was dried over anhydrous sodium sulfate, filtered, concentrated to remove the solvent, and dried in vacuo to give the target compound II .

More specific examples (compound of Example 112 as described below) are as follows:

To a 25 mL flask was added 27 mg of compound 9-1 . 3 mL of methanol was added as solvent, followed by 3 mL of saturated sodium bicarbonate solution. The mixture was stirred and reacted at room temperature for 4 hours. After the reaction was completed, the reaction mixture was concentrated to remove a portion of methanol and extracted with dichloromethane. The resulting organic layer was dried over anhydrous sodium sulfate, filtered, concentrated to remove the solvent, and dried in vacuo to give 20.5 mg of target compound 9-2 .

Test Example 1: Analysis of In Vitro Inhibitory Activity on CDK Kinase

In an in vitro cell-free kinase activity assay, the half inhibitory concentration (IC ₅₀ ) of a compound of the present invention against CDK6/CyclinD1 (trade name: Carna, product number 04-114) was detected as follows. It should be noted that the assay can be performed in a similar way. The test steps of this test example are as follows.

CDK6/CyclinD1 kinase activity was determined using a time-resolved fluorescence resonance energy transfer (TR-FRET) method. Measurements were performed in a reaction volume of 20 μL using a 384-well assay plate. Kinases, inhibitors, ATP (concentration equivalent to K _m for CDK6/CyclinD1 kinase, trade name: Sigma, product no. A7699) and polypeptide substrates (concentration equivalent to K _m for CDK6/CyclinD1 kinase, trade name: Cisbio, product number 64CUS000C01B) 20 mM HEPES (pH 7.5), 1 mM EGTA, 10 mM MgCl ₂ , 0.02% Briji35, 0.02 mg/ml BSA, 0.1 mM Na ₃ VO ₄ , 2 mM DTT, and 1% DMSO (pH 7.5). Incubated for 1.5 hours in buffer. Reaction was carried out by addition of a mixture of peptide antibody (trade name: Cisbio, product number 64CUSKAY) and Sa-XL665 (trade name: Cisbio, product number 610SAXLB) diluted in HTRF KinEASE detection buffer (trade name: Cisbio, product number 62SDBRDF, containing EDTA) stopped. After the mixture was incubated at room temperature for 1 hour, the plate was read. The TR-FRET signal was analyzed on a multimode plate reader (EnVision ^™ , multi-function microplate detector, PerkinElmer) at an excitation wavelength (λ _Ex ) of 330 nm and detection wavelengths (λ _Em ) of 615 and 665 nm. measured. Activity was determined by the ratio of fluorescence at 665 nm to fluorescence at 615 nm. For each compound, the IC ₅₀ was determined by measuring the enzymatic activity of different concentrations of the compound. The negative control contained no inhibitor. Two non-enzymatic controls were used to determine baseline fluorescence levels. IC ₅₀ was obtained by fitting using GraphPad 6.02 software.

Each of the exemplary compounds and the reference example compounds was prepared according to the scheme described above, and details are shown in Table 1. For each compound, 8-10 concentrations were prepared and formulated in DMSO. The stock concentration was 200 times the working concentration. For example, if the working concentration is 100 nM, the corresponding concentration of the formulated stock solution is 20 μM.

In the assay of inhibitory activity in vitro against CDK kinase, IC ₅₀ values of each exemplary compound and reference compound were determined. IC ₅₀ values are given according to the interval of IC ₅₀ values, where "+++" indicates IC ₅₀ < 100 nM; "++" indicates 100 nM ≤ IC ₅₀ < 1000 nM; "+" represents 1000 nM ≤ IC ₅₀ < 3000 nM; and "-" stands for "more than 3000 nM".

As can be seen from Table 1, exemplary compounds falling within the protection scope of the present invention are well capable of inhibiting the activity of CDK6 kinase.

Test Example 2: Cell viability assay

In vitro cell viability assays were performed using some exemplary compounds according to the following steps:

1. After recovery, MCF-7 cells (obtained from Shanghai Cell Bank (Chinese Academy of Sciences)) were cultured at 37° C., 5% CO ₂ and 95% humidity.

2. Eight compound solutions of different concentrations were prepared for each exemplary compound. 50 μL of each concentration of compound solution was added to a 96-well black plate.

3. The cell concentration was adjusted to about 30,000 cells/mL. 100 μL of the cell suspension was added to a 96-well plate and mixed well to a final cell density of about 3,000 cells/well.

4. 96 well plates were incubated at 37° C., 5% CO ₂ and 95% humidity for 168 hours, ie 7 days.

5. Cell viability was measured by CellTiter-Glo ^® Luminescent Cell Viability Assay (Promega, Cat#G7572). Fluorescence data were read on a microplate reader (EnVision ^™ , Multifunction Microplate Detector, PerkinElmer).

6. The obtained fluorescence data were analyzed using GraphPad Prism software. Cell viability was calculated for the exemplary compound at each concentration, and the IC ₅₀ interval of each exemplary compound was determined.

IC ₅₀ values of exemplary compounds are listed in Table 2 below, where "+++" indicates IC ₅₀ < 500 nM; "++" indicates 500 nM ≤ IC ₅₀ < 2500 nM; "+" indicates 2500 nM ≤ IC ₅₀ < 10000 nM; "-" indicates greater than 10000 nM.

As can be seen from the above table, the compound of the present invention can achieve an excellent inhibitory effect on MCF-7 cells.

The examples and implementations described herein are for illustrative purposes only, and in light of this, various modifications or changes will suggest to those skilled in the art, and are within the spirit and scope of this application and the scope of the appended claims. understood to be included. All publications, patents and patent applications cited herein are hereby incorporated by reference in their entirety for all purposes.

Claims (10)

A compound of formula (I) or a pharmaceutically acceptable salt thereof:

during the meal,
Q is an optionally substituted 6- to 18-membered arylene group or an optionally substituted 5- to 18-membered heteroarylene group, wherein when substituted, the substituents are halo, _hydroxy , C _1-6 alkyl, selected from the group consisting of C _1-6 alkoxy, and halo _- C _{1-6 alkyl} ;
R ₁ is an optionally substituted 3- to 8-membered heterocyclyl group, an optionally substituted 6- to 14-membered fused heterocyclyl group, or an optionally substituted 6- to 12-membered spiro heterocyclyl group wherein, when substituted, the substituent is selected from the group consisting of halo, hydroxy, C _1-6 alkyl, C _1-6 alkoxy, and halo-C _1-6 alkyl;
R ₂ is H, halo, optionally substituted 3- to 10-membered cycloalkenyl group, optionally substituted 3- to 10-membered heterocycloalkenyl group, optionally substituted 3- to 10-membered cycloalkyl group, an optionally substituted 3- to 10-membered heterocycloalkyl group, an optionally substituted 6- to 18-membered aryl group, or an optionally substituted 5- to 18-membered heteroaryl group, wherein substituted , the substituents are selected from the group consisting of halo, _hydroxy , C _{1-6 alkyl} , C _1-6 alkoxy, halo _- C _{1-6 alkyl} , C _1-6 alkoxy _- C _1-6 alkoxy, and oxo;
R ₃ is H, CN, —C(=O)—NR ₄ R ₅ , an optionally substituted 6- to 18-membered aryl group, an optionally substituted 5- to 18-membered heteroaryl group, or optionally substituted a 5- to 8-membered lactam group wherein, when substituted, the substituent is selected from the group consisting of halo, _hydroxy , C _{1-6 alkyl} , C _1-6 alkoxy, and halo _- C _1-6 alkyl;
R ₄ and R ₅ are each independently methyl or ethyl; And
when R ₃ is —C(=O)—NR ₄ R ₅ , then R ₂ is an optionally substituted 3- to 10-membered cycloalkenyl group, an optionally substituted 3- to 10-membered heterocycloalkenyl group , an optionally substituted 3- to 10-membered heterocycloalkyl group linked at the N atom to a non-R ₂ structure of Formula (I), or an optionally substituted 6- to 18-membered aryl group, wherein substituted , the substituent is selected from the group consisting of halo, _hydroxy , C _1-6 alkyl, C _1-6 alkoxy, halo _- C _{1-6 alkyl} , C _1-6 alkoxy _- C _1-6 alkoxy, and oxo. According to claim 1,
Q is optionally substituted phenylene or optionally substituted pyridinylene, wherein when substituted, the substituents are halo, _hydroxy , C _{1-6 alkyl} , C _1-6 alkoxy, and halo _- C _1-6 alkyl. is selected from the group consisting of;
R ₁ is an optionally substituted 3- to 8-membered heterocyclyl group, an optionally substituted 6- to 14-membered fused heterocyclyl group, or an optionally substituted 6- to 12-membered spiro heterocyclyl group wherein when substituted, the substituent is selected from C _{1-6 alkyl} ;
R ₂ is a halogen atom, optionally substituted cyclopentenyl, optionally substituted cyclohexenyl, optionally substituted cyclopentyl, optionally substituted cyclohexyl, optionally substituted oxacyclohexenyl, optionally substituted aza cyclohexenyl, optionally substituted oxolanyl, optionally substituted azacyclopentyl, optionally substituted oxacyclohexyl, optionally substituted azacyclohexyl, optionally substituted phenyl, optionally substituted naphthyl, optionally is selected from the group consisting of pyridinyl substituted with , optionally substituted thienyl, optionally substituted pyrazolyl, optionally substituted oxazolyl, optionally substituted isoxazolyl, and optionally substituted quinolyl, wherein When substituted, the substituent is selected from the group consisting of halo, _hydroxy , C _1-6 alkyl, C _1-6 alkoxy, halo _- C _{1-6 alkyl} , and oxo;
R ₃ is H, -CN, -C(=O)-NR ₄ R ₅ , optionally substituted phenyl, naphthyl, pyrazolyl, pyridinyl, thienyl, oxazolyl, isoxazolyl, pyrimidinyl, imine Dazolyl, pyrrolyl,

or

wherein, when substituted, the substituent is selected from the group consisting of halo, _hydroxy , C _{1-6 alkyl} , C _1-6 alkoxy and halo _- C _1-6 alkyl; And
R ₄ and R ₅ are both methyl; or a pharmaceutically acceptable salt thereof. 3. The method according to claim 1 or 2, wherein Q is phenylene or

Phosphorus, a compound, or a pharmaceutically acceptable salt thereof. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, wherein R 1 is an optionally substituted group as follows, and when substituted, the substituent is selected from C _{1-6 alkyl} . :

5. The compound or pharmaceutically acceptable salt thereof according to claim 4, wherein C _1-6 alkyl is _methyl or ethyl. 6 . The compound or pharmaceutically thereof according to claim 1 , wherein R ₂ is halo or an optionally substituted group, wherein when substituted, the substituent is selected from C _{1-6 alkyl} . Acceptable salts as:

The compound or pharmaceutically acceptable salt thereof according to claim 1, which is selected from the group consisting of:

and

8. The compound according to any one of claims 1 to 7, for inhibiting the activity of CDK4/6 kinase. 9. A pharmaceutical composition comprising a therapeutically effective amount of a compound of any one of claims 1 to 8, and a pharmaceutically acceptable carrier or excipient. The use of the compound of any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment and/or prevention of a cancer-related disease mediated by CDK4/6 kinase, wherein the cancer-related Diseases include brain tumor, lung cancer, squamous cell carcinoma, bladder cancer, stomach cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, rectal cancer, liver cancer, kidney cancer, esophageal adenocarcinoma, esophageal squamous cell carcinoma, prostate cancer, female reproductive tract cancer, carcinoma in situ, lymphoma, neurofibroma, thyroid cancer, bone cancer, skin cancer, brain cancer, colon cancer, testicular cancer, gastrointestinal stromal tumor, prostate tumor, mast cell tumor, multiple myeloma, melanoma, glioma, or a use selected from breeding.